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Head and Neck Surgery

Macrophage Toll-Like Receptor Activation Modulates Oral SCC Invasion

Sunday, October 1, 2023
1:42 PM - 1:48 PM East Coast USA Time
Location: 201AB

    Presenting Author(s)

  • OR

    Olivia L. Ricks, RN BSN

    1Heersink School of Medicine, University of Alabama at Birmingham
    Birmingham, AL, USA

    • Co-Author(s)

  • AM

    Ameer Mansur, BS

    2Department of Biomedical Engineering, University of Alabama at Birmingham, AL, USA

  • JP

    Juhi J. Patel, BS

    Researcher II
    4Department of Otolaryngology – Head and Neck Surgery, University of Alabama at Birmingham
    Madison, AL, United States

  • ZE

    Zoë E. Evans, BS

    5Cancer Biology Program, University of Alabama at Birmingham,, AL, USA

  • GF

    Gabriela S. Fonseca, BS

    1Heersink School of Medicine, University of Alabama at Birmingham, AL, USA

  • AS

    Anna G. Sorace, PhD

    3Department of Radiology, University of Alabama at Birmingham, AL, USA

    • Senior Author(s)

  • CT

    Carissa M. Thomas, MD, PhD, FACS

    Assistant Professor
    University of Alabama at Birmingham
    Birmingham, AL, United States

Introduction: Select oral bacteria may activate macrophages in the tumor microenvironment (TME) and influence oral squamous cell carcinoma (OSCC) invasion. Our objective is to determine if toll-like receptor (TLR) or bacteria conditioned media (BCM) activation of macrophages can modulate OSCC invasion.

Methods: THP-1 monocytes were treated with phorbol-12-myristate-13-acetate (PMA) to differentiate into macrophages and then activated with TLR agonists or BCM (Streptococcus sanguinis strain KTH3 and Fusobacterium nucleatum strain nucleatum grown to stationary phase and filter sterilized). Macrophages stimulated by TLR agonists were treated with 100ng/mL lipopolysaccharide (LPS) or 100ng/mL Pam3CSK4 (PCK), for 24, 48, and 72 hours. Macrophages stimulated by BCM were treated with 100uL BCM for 6, 12, 24, 36, or 48 hours. SCC-1 (floor of mouth) and OSC-19 (tongue) were cultured with the secreted factors from TLR or BCM-activated macrophages in a Matrigel invasion assay system (Corning, Tewksbury, MA) for 24 hours (n=3). Crystal violet staining was performed, and membranes were imaged using EVOS7000. Quantification of invasion was determined using a custom MATLAB program, blinded cell counting, and acetic acid elution of crystal violet and absorbance measurements.

Results: SCC-1 and OSC-19 have minimal baseline invasiveness. Secreted factors from TLR4-activated macrophages (24, 48, 72 hours) increased invasion of OSC-19 cells (4.4%, 10.3% (p=0.0216), 7.2% and 0.832nm, 2.119nm (p=0.0139), 1.502nm respectively). Secreted factors from TLR2-activated macrophages (24, 72 hours) increased invasion of OSC-19 cells (5.5%, 12.6% (p=0.006) and 0.933nm, 1.595nm respectively). Invasion of SCC-1 cells was not affected by secreted factors from TLR-activated macrophages. Activation of macrophages with secreted factors from S. sanguinis KTH3 and F. nucleatum did not increase invasion of SCC-1 or OSC-19 cells.

Conclusions: Activation of macrophages with TLR agonists increases the invasiveness of OSC-19. Bacterial interactions with immune cells of the TME may influence cancer cell invasion.